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The effect on the transcriptome of Anemone coronaria following infection with rust (Tranzschelia discolor).

Laura M, Borghi C, Bobbio V, Allavena A - PLoS ONE (2015)

Bottom Line: Unigene annotation showed that 50.5% of the predicted translation products shared significant homology with protein sequences in GenBank.Only one R gene (PGIP) was up-regulated.The DTA genes identified here should provide a basis for understanding the A. coronaria/T. discolor interaction and leads for biotechnology-based disease resistance breeding.

View Article: PubMed Central - PubMed

Affiliation: CRA-Unità di Ricerca per la Floricoltura e le Specie Ornamentali, Corso Inglesi 508, 18038 Sanremo (IM), Italy.

ABSTRACT
In order to understand plant/pathogen interaction, the transcriptome of uninfected (1S) and infected (2I) plant was sequenced at 3'end by the GS FLX 454 platform. De novo assembly of high-quality reads generated 27,231 contigs leaving 37,191 singletons in the 1S and 38,393 in the 2I libraries. ESTcalc tool suggested that 71% of the transcriptome had been captured, with 99% of the genes present being represented by at least one read. Unigene annotation showed that 50.5% of the predicted translation products shared significant homology with protein sequences in GenBank. In all 253 differential transcript abundance (DTAs) were in higher abundance and 52 in lower abundance in the 2I library. 128 higher abundance DTA genes were of fungal origin and 49 were clearly plant sequences. A tBLASTn-based search of the sequences using as query the full length predicted polypeptide product of 50 R genes identified 16 R gene products. Only one R gene (PGIP) was up-regulated. The response of the plant to fungal invasion included the up-regulation of several pathogenesis related protein (PR) genes involved in JA signaling and other genes associated with defense response and down regulation of cell wall associated genes, non-race-specific disease resistance1 (NDR1) and other genes like myb, presqualene diphosphate phosphatase (PSDPase), a UDP-glycosyltransferase 74E2-like (UGT). The DTA genes identified here should provide a basis for understanding the A. coronaria/T. discolor interaction and leads for biotechnology-based disease resistance breeding.

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Phylogenetic tree and aligment of thaumatin-like protein.(A)Anemone coronaria thaumatin-like protein (contig 13789) cluster with Puccinia graminis and Melampsora larici-populina proteins. TLPs of no-rust fungal taxa cluster in a separate group. Bootstrap values are indicated in relevant nodes. Arabidopsis thaliana TLP was used as out-group. (B) Amino acid sequence alignment of five rust TLPs. The conserved 16 cystein residues are highlighted in the boxes.
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pone.0118565.g003: Phylogenetic tree and aligment of thaumatin-like protein.(A)Anemone coronaria thaumatin-like protein (contig 13789) cluster with Puccinia graminis and Melampsora larici-populina proteins. TLPs of no-rust fungal taxa cluster in a separate group. Bootstrap values are indicated in relevant nodes. Arabidopsis thaliana TLP was used as out-group. (B) Amino acid sequence alignment of five rust TLPs. The conserved 16 cystein residues are highlighted in the boxes.

Mentions: Among the likely fungal sequences, 118 had homologs in either P. graminis f. sp. tritici or Melampsora larici-populina and 78 were associated with a likely function (S2 Table). Of the 30 genes encoding ribosomal proteins (RPs), 26 were likely to have been of fungal origin, reflecting the active protein synthesis exhibited by fungi during the early phase of infection [48]. Genes hydrolytic enzymes acting on plant biopolymers (cellulase), proteinase (subtilase-type proteinase psp3, vacuolar protease A, proteasome subunit 1) and several carbohydrate-active enzymes (glycoside hydrolase, glyceraldehyde-3-phosphate dehydrogenase, enolase, glucose-repressible protein) were well represented, as would be predicted since the invading fungus penetrates the host cells by degrading enzymes [49,50]. Apart from these, a fungal chitinase gene was recognized; this enzyme is used to remodel fungal cell wall during infection, either to promote hyphal invasion and/or to avoid recognition by the host’s defense system [51]. Other strongly represented fungal genes encoded histones, an argonaute-like protein, thiamine synthesis, a mitochondrial thiazole synthesis enzyme and the ubiquitin-conjugating enzyme E2, as was also the case during the infection by rust of both Populus sp. and wheat [49]. P. triticina genes encoding a thiamine synthesis protein and a cyclophilin are also induced in planta, as reported by Thara et al. [48]. The first protein is a cofactor controlling the activity of several enzymes involved in the central carbon metabolism [52], whereas cyclophilin is involved in a wide variety of cellular processes, including the response to abiotic stress, the control of cell cycling, the regulation of calcium signaling and control of transcriptional repression [53–55]. The transcription of a gene encoding a thaumatin-like protein (TLP) may reflect the fungus’ attempt to interfere with the host’s defense signaling apparatus [56,57]; the presence of this protein has been noted in the Melampsora secretome [58,59]. The phylogenetic tree of TLP resolved the entries into two major branches. One includes rust fungal proteins only (P. graminis and M. larici-populina and T. discolor), the other groups proteins of all other fungal taxa. T. discolor TLP is well separated from P. graminis and M. larici-populina proteins with a bootstrap of 99% (Fig. 3A). Amino acid sequence of T. discolor TLP shares with rust TPLs the 16 conserved cysteine residues that characterize the large type TLPs [60].


The effect on the transcriptome of Anemone coronaria following infection with rust (Tranzschelia discolor).

Laura M, Borghi C, Bobbio V, Allavena A - PLoS ONE (2015)

Phylogenetic tree and aligment of thaumatin-like protein.(A)Anemone coronaria thaumatin-like protein (contig 13789) cluster with Puccinia graminis and Melampsora larici-populina proteins. TLPs of no-rust fungal taxa cluster in a separate group. Bootstrap values are indicated in relevant nodes. Arabidopsis thaliana TLP was used as out-group. (B) Amino acid sequence alignment of five rust TLPs. The conserved 16 cystein residues are highlighted in the boxes.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4359109&req=5

pone.0118565.g003: Phylogenetic tree and aligment of thaumatin-like protein.(A)Anemone coronaria thaumatin-like protein (contig 13789) cluster with Puccinia graminis and Melampsora larici-populina proteins. TLPs of no-rust fungal taxa cluster in a separate group. Bootstrap values are indicated in relevant nodes. Arabidopsis thaliana TLP was used as out-group. (B) Amino acid sequence alignment of five rust TLPs. The conserved 16 cystein residues are highlighted in the boxes.
Mentions: Among the likely fungal sequences, 118 had homologs in either P. graminis f. sp. tritici or Melampsora larici-populina and 78 were associated with a likely function (S2 Table). Of the 30 genes encoding ribosomal proteins (RPs), 26 were likely to have been of fungal origin, reflecting the active protein synthesis exhibited by fungi during the early phase of infection [48]. Genes hydrolytic enzymes acting on plant biopolymers (cellulase), proteinase (subtilase-type proteinase psp3, vacuolar protease A, proteasome subunit 1) and several carbohydrate-active enzymes (glycoside hydrolase, glyceraldehyde-3-phosphate dehydrogenase, enolase, glucose-repressible protein) were well represented, as would be predicted since the invading fungus penetrates the host cells by degrading enzymes [49,50]. Apart from these, a fungal chitinase gene was recognized; this enzyme is used to remodel fungal cell wall during infection, either to promote hyphal invasion and/or to avoid recognition by the host’s defense system [51]. Other strongly represented fungal genes encoded histones, an argonaute-like protein, thiamine synthesis, a mitochondrial thiazole synthesis enzyme and the ubiquitin-conjugating enzyme E2, as was also the case during the infection by rust of both Populus sp. and wheat [49]. P. triticina genes encoding a thiamine synthesis protein and a cyclophilin are also induced in planta, as reported by Thara et al. [48]. The first protein is a cofactor controlling the activity of several enzymes involved in the central carbon metabolism [52], whereas cyclophilin is involved in a wide variety of cellular processes, including the response to abiotic stress, the control of cell cycling, the regulation of calcium signaling and control of transcriptional repression [53–55]. The transcription of a gene encoding a thaumatin-like protein (TLP) may reflect the fungus’ attempt to interfere with the host’s defense signaling apparatus [56,57]; the presence of this protein has been noted in the Melampsora secretome [58,59]. The phylogenetic tree of TLP resolved the entries into two major branches. One includes rust fungal proteins only (P. graminis and M. larici-populina and T. discolor), the other groups proteins of all other fungal taxa. T. discolor TLP is well separated from P. graminis and M. larici-populina proteins with a bootstrap of 99% (Fig. 3A). Amino acid sequence of T. discolor TLP shares with rust TPLs the 16 conserved cysteine residues that characterize the large type TLPs [60].

Bottom Line: Unigene annotation showed that 50.5% of the predicted translation products shared significant homology with protein sequences in GenBank.Only one R gene (PGIP) was up-regulated.The DTA genes identified here should provide a basis for understanding the A. coronaria/T. discolor interaction and leads for biotechnology-based disease resistance breeding.

View Article: PubMed Central - PubMed

Affiliation: CRA-Unità di Ricerca per la Floricoltura e le Specie Ornamentali, Corso Inglesi 508, 18038 Sanremo (IM), Italy.

ABSTRACT
In order to understand plant/pathogen interaction, the transcriptome of uninfected (1S) and infected (2I) plant was sequenced at 3'end by the GS FLX 454 platform. De novo assembly of high-quality reads generated 27,231 contigs leaving 37,191 singletons in the 1S and 38,393 in the 2I libraries. ESTcalc tool suggested that 71% of the transcriptome had been captured, with 99% of the genes present being represented by at least one read. Unigene annotation showed that 50.5% of the predicted translation products shared significant homology with protein sequences in GenBank. In all 253 differential transcript abundance (DTAs) were in higher abundance and 52 in lower abundance in the 2I library. 128 higher abundance DTA genes were of fungal origin and 49 were clearly plant sequences. A tBLASTn-based search of the sequences using as query the full length predicted polypeptide product of 50 R genes identified 16 R gene products. Only one R gene (PGIP) was up-regulated. The response of the plant to fungal invasion included the up-regulation of several pathogenesis related protein (PR) genes involved in JA signaling and other genes associated with defense response and down regulation of cell wall associated genes, non-race-specific disease resistance1 (NDR1) and other genes like myb, presqualene diphosphate phosphatase (PSDPase), a UDP-glycosyltransferase 74E2-like (UGT). The DTA genes identified here should provide a basis for understanding the A. coronaria/T. discolor interaction and leads for biotechnology-based disease resistance breeding.

Show MeSH
Related in: MedlinePlus